Difference between revisions of "Part:BBa K2145104:Experience"

 
 
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
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how you used this part and how it worked out.
 
  
 
===Applications of BBa_K2145104===
 
===Applications of BBa_K2145104===
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[[File:Splates95102.png]]<br>
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Qualitatively, we noticed that none of the colonies containing this plasmid showed BOTH visible RFP and visible GFP -- each colony was either red, green, or neither, but never red and green (see GG97 above). Fluorescence was observed even without the addition of inducers (IPTG and ATc), even though the reporters should be under IPTG and ATc control. We attribute this to insufficient repression by LacI and TetR.
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[[File:Alverno_ca_97.png]]
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[[File:Alverno_ca_r_97.png]]
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[[File:Alverno_ca_g_97.png]]
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Shown above are OD600 measurements, RFP fluorescence, and GFP fluorescence, respectively, for three clones containing this part in liquid culture. Again, we emphasize that only RFP OR GFP are expressed, but never both. We believe this may be due to supercoiling -- when one gene expresses, it creates supercoils that shut down the other gene.
  
 
===User Reviews===
 
===User Reviews===
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Enter the review inofrmation here.
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This part contains two fluorescent protein coding sites (for RFP and GFP) and was assembled using Golden Gate Assembly. To make plasmid, parts were taken from iGEM Registry (BBa_J04450 (RFP) and BBa_I13522 (GFP)) and assembled to make new part with a 500bp spacer in between.
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Direction of GFP: Reverse, away from the spacer; Direction of RFP: Forward, away from the spacer
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Latest revision as of 00:31, 31 October 2016


Applications of BBa_K2145104

Splates95102.png
Qualitatively, we noticed that none of the colonies containing this plasmid showed BOTH visible RFP and visible GFP -- each colony was either red, green, or neither, but never red and green (see GG97 above). Fluorescence was observed even without the addition of inducers (IPTG and ATc), even though the reporters should be under IPTG and ATc control. We attribute this to insufficient repression by LacI and TetR.

Alverno ca 97.png Alverno ca r 97.png Alverno ca g 97.png

Shown above are OD600 measurements, RFP fluorescence, and GFP fluorescence, respectively, for three clones containing this part in liquid culture. Again, we emphasize that only RFP OR GFP are expressed, but never both. We believe this may be due to supercoiling -- when one gene expresses, it creates supercoils that shut down the other gene.

User Reviews

UNIQ6ae085e7f3b32c4e-partinfo-00000000-QINU UNIQ6ae085e7f3b32c4e-partinfo-00000001-QINU